Microarray dispensing with real-time verification and inspection

ABSTRACT

A microarrayer for spotting solution onto a receiving surface in an automated microarray dispensing device. Elements of the present invention include: at least one dispense head for spotting the receiving surface, at least one light source capable of illuminating the receiving surface, at least one camera operating in conjunction with the at least one light source. The at least one camera is capable of acquiring and transmitting surface image data to a computer. The computer is programmed to receive the surface image data and analyze it. The computer will then generate post analysis data based on the analysis of the surface image data. The post analysis data is available for improving the spotting of the solution onto the receiving surface. In a preferred embodiment, the surface image data includes information relating to receiving surface alignment, information relating to spot quality, and receiving surface identification information. In a preferred embodiment, the analysis of the information relating to receiving surface alignment enables the computer to make automatic adjustments to the relative positions of the at least one dispense head and the receiving surface to increase the accuracy of the spotting. In a preferred embodiment, the analysis of the information relating to spot quality identifies a spot as pass or fail. An operator is then able to rework the spot. In a preferred embodiment, the analysis of the receiving surface identification information enables the computer to track each receiving surface. In a preferred embodiment the receiving surface is a plurality of slides.

[0001] The present invention relates to automated microarray dispensingdevices, more specifically it relates to automated microarray dispensingdevices with automated quality inspection capability. This applicationis a continuation in part application of U.S. patent application SerialNo. 09/611,256 filed Jul. 6, 2000, which is incorporated by referenceherein.

BACKGROUND OF THE INVENTION

[0002] Microarrays, also known as biochips, have recently becomeimportant in the study of genomics. The use of a microarray involveslaying down an ordered array of genetic elements onto a solid substratesuch as a slide. Depending on the application, a microarray may consistof genomic DNA, reverse-transcribed cDNA, or smaller chains ofoligonucleotides as well as any preparatory substrates. The microarrayis useful because it allows genetic analysis to take place on amassively parallel scale, wherein thousands of genes and markers can bescored in one experiment.

[0003] A microarrayer, also known as a DNA array printer, is ahigh-capacity system used to print a microarray onto slides. Typically,a microarrayer is a specially built robotic platform designed totransfer solutions from the well of some type of microplate onto anothersurface for analysis. This process of depositing the liquid spot ontothe slide is known as “spotting”.

[0004] Recently, microarrayers have become extremely popular inlaboratories because they add to the efficient productivity of thelaboratory to be able to print samples onto slides accurately andrapidly. Affymetrix, Inc., with offices in Santa Clara, California,makes an automated arrayer called the 417 ARRAYER (Part No. 0-0001 andPart No. 0-0009). BioRobotics, with offices in Boston, Massachusetts,produces two versions of an automated arrayer called the MICROGRID andMICROGRID II. GeneMachines, with offices in Menlo Park, Calif. makes anarrayer called the OMNIGRID (Model No. OGR-02). Packard InstrumentCompany with offices in Meriden, Connecticut makes an automated arrayercalled the BIOCHIP ARRAYER.

[0005] Although there are some differences between each of the abovelisted microarrayers, they are all similar in that they each spotmicroarrays in an automated fashion. However, there are significantproblems with the prior art devices that detracts from their efficientoperation.

[0006] A first problem arises due to the fact that as blank slides arecycled through prior art microarrayers, they can become askew orpositioned improperly underneath dispensing tips. This problem resultsin spots being positioned improperly on the slides. A second problem canarise even if the slide is positioned correctly under the dispensingtips. It is possible for the spot to be deposited in the correctposition, but be of poor quality and therefore useless as far asexperimentation purposes.

[0007] Up to now, the only way to deal with these problems was to have ahuman operator visually monitor and inspect the microarrayer during itsoperation or inspect the samples after they come off the machine. Thissolution is an unacceptable waste of human effort. The BIOCHIP ARRAYERmade by Packard Instrument Company has attempted to deal with theproblem of monitoring the spotting process. However, it has only limitedverification functionality with its integrated camera. This means thatit verifies whether or not a spot has been dispensed, without anyquality inspection to analyze whether that spot was good or bad.

[0008] What is needed is a better microarrayer with automated qualityinspection capability.

SUMMARY OF THE INVENTION

[0009] The present invention provides a microarrayer for spottingsolution onto a receiving surface in an automated microarray dispensingdevice. Elements of the present invention include: at least one dispensehead for spotting the receiving surface, at least one light sourcecapable of illuminating the receiving surface, at least one cameraoperating in conjunction with the at least one light source. The atleast one camera is capable of acquiring and transmitting surface imagedata to a computer. The computer is programmed to receive the surfaceimage data and analyze it. The computer will then generate post analysisdata based on the analysis of the surface image data. The post analysisdata is available for improving the spotting of the solution onto thereceiving surface. In a preferred embodiment, the surface image dataincludes information relating to receiving surface alignment,information relating to spot quality, and receiving surfaceidentification information. In a preferred embodiment, the analysis ofthe information relating to receiving surface alignment enables thecomputer to make automatic adjustments to the relative positions of theat least one dispense head and the receiving surface to increase theaccuracy of the spotting. In a preferred embodiment, the analysis of theinformation relating to spot quality identifies a spot as pass or fail.An operator is then able to rework the spot. In a preferred embodiment,the analysis of the receiving surface identification information enablesthe computer to track each receiving surface. In a preferred embodimentthe receiving surface is a plurality of slides.

BRIEF DESCRIPTION OF THE DRAWINGS

[0010]FIG. 1 shows the major components of a preferred embodiment of thepresent invention.

[0011]FIG. 2 shows slides fifty slides located on 5 locating platesafter the slides have been spotted.

[0012] FIGS. 3-32 illustrate the sequence of operation of a preferredembodiment of the present invention.

[0013]FIG. 33 shows the major components of a preferred embodiment ofthe present invention.

[0014]FIG. 34A shows the rework dispense head in the up position.

[0015]FIG. 34B shows the rework dispense head in the down position.

[0016]FIG. 34C shows a slide with a 2D bar code.

[0017]FIGS. 35A and 35B shows dispense tips attached to dispense heads.

[0018]FIG. 36 shows a preferred embodiment of the present inventionmounted on a vibration isolated base.

[0019]FIG. 37 shows the major components of a preferred embodiment ofthe present invention.

[0020]FIGS. 38A and 38B show a flowchart for the programming of apreferred embodiment of the present invention.

[0021]FIGS. 39A and 39B show the reworking of a slide for a preferredembodiment of the present invention.

[0022]FIG. 40 shows another preferred embodiment of the presentinvention.

[0023]FIG. 41 shows another preferred embodiment of the presentinvention.

[0024]FIG. 42 shows another preferred embodiment of the presentinvention.

[0025]FIG. 43 shows another preferred embodiment of the presentinvention.

[0026]FIG. 44 shows a dispense head over a microplate.

[0027] FIGS. 45A-45C show a pre-spot slide.

[0028]FIG. 46 shows a dispense head over a slide.

[0029]FIG. 47 shows a preferred incubator.

[0030]FIG. 48 shows robotic plate handling.

[0031] FIGS. 49A-49B show a microplate.

[0032] FIGS. 49C-49D show a dispense head over a microplate.

[0033]FIGS. 50 and 51 show a preferred microplate and dispense head.

[0034]FIG. 52A shows another preferred embodiment of the presentinvention.

[0035]FIG. 52B shows microplates in storage racks.

[0036]FIG. 53 shows spots deposited on a preferred slide.

[0037] FIGS. 54-58 show a preferred monitor screen display.

[0038]FIG. 59 shows spot inspection data.

[0039]FIG. 60 shows dispense pin spot inspection analysis.

[0040]FIG. 61 shows a calibration target.

[0041]FIG. 62 shows another preferred monitor screen display.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

[0042] A detailed description of a preferred embodiment of the presentinvention can be described by reference to FIGS. 1-38B. During theoperation of the present invention, solution from reservoir plate 5 isautomatically deposited onto an array of fifty blank slides 4A1-4E10located on locating plates 4A-4E (see FIG. 1). An operator is able toselect via a computer interface whether dispense tip 42 (locatedunderneath dispense head 40) or a 4×6 array of dispense tips 7 (locatedunderneath dispense head 6) will be used to make the deposits ontoslides 4A1-4E10. In the preferred embodiment, dispense tips 7 and 42 arequill type dispense tips. Locating plates 4A-4E are mounted on linearactuator 15 so that they can move along the x-axis. Linear actuator 26is mounted on linear actuator 21 so that linear actuator 26 can movealong the y-axis. Dispense heads 6 and 40 are mounted on linear actuator26 so that they can move along the z-axis. Camera 12 with strobe light13 is focused so as to permit recording of the deposition process andfunctions to permit verification of slide identification information andto permit verification of proper deposition of solution on the slides.Periodically, during the cycle, the dispense tips are cleaned in soniccleaner 9, rinsed in rinse fountain 10, and then dried in vacuummanifold 11. After the solution has been deposited onto the slides, theoperator can retrieve locating plates 4A-4E containing slides 4A1-4E10from the position shown in FIG. 2.

Sequence of Operation of a Preferred Embodiment

[0043] FIGS. 3-32 illustrate the sequence of operation of a preferredembodiment of the present invention.

[0044] In a preferred embodiment of the present invention, the operationof the components is controlled by PC control system 300, as shown inFIG. 37. FIGS. 38A-38E show a flowchart representing preferredprogramming of PC control system 300 and corresponds to the sequenceillustrated in FIGS. 3-32.

[0045] As shown in FIG. 3, an operator places five locating plates 4A-4Eeach having ten clean, blank slides 4A1-4E10 on platform 2. In apreferred embodiment of the present invention, slides 4A1-4E10 are madeby Sequonem with offices in San Diego, Calif. A preferred slide is shownin FIG. 16B. It has ninety-six etched dispense positions 60 and has itsown unique 2D bar code 65 for identification purposes.

[0046] As shown in FIG. 4, linear actuator 15 moves platform 2 so thatslide 4A1 is underneath the dispense head 6. Dispense head 6 ispositioned directly above slide 4A1. Using camera 12 and the strobelight 13, an image is acquired of slide 4A1. The camera reads the barcode and inspects the positioning and alignment of slide 4A1 on locatingplate 3A. The software then analyzes the position data and stores theinformation. The information stored and will be used later to adjust thepositions of slide 4A1 and dispense head 6 to ensure accurate placementof the solution on the slide.

[0047] After camera 12 has acquired the image of slide 4A1, linearactuator 26 is moved via linear actuator 21 to the position shown inFIG. 5 so that dispense head 6 is directly above slide 4A2. Using camera12 and the strobe light 13, an image is acquired of slide 4A2. As withslide 4A1, camera reads the bar code and inspects the positioning andalignment of slide 4A2 on locating plate 4A. The software then analyzesthe position data and stores the information. The information stored andwill be used later to adjust the positions of slide 4A2 and dispensehead 6 to ensure accurate placement of the solution on the slide.

[0048] Linear actuator 26 is then moved via linear actuator 21 to theposition shown in FIG. 6 so that dispense head 6 is directly above soniccleaner 9.

[0049] As shown in FIG. 7, mounting plate 25 moves downward via linearactuator 26 so that dispense tips 7 are dipped in sonic cleaner 9 for aprogrammable time period while the cleaner is turned on. When finished,mounting plate 25 moves upward as shown in FIG. 8.

[0050] Then, as shown in FIG. 9, platform 31 moves to the left viapneumatic slide 30, thereby moving rinse fountain 10 and vacuum manifold11 to the left. Linear actuator 26 is moved via linear actuator 21 backso that it is directly above rinse fountain 10.

[0051] As shown in FIG. 10, mounting plate 25 moves downward via linearactuator 26 so that tips 7 are dipped in rinse fountain 10 for aprogrammable time period while the fountain is turned on.

[0052] After rinsing, mounting plate 25 moves upward via linear actuator26, as shown in FIG. 11. Platform 31 moves to the right via pneumaticslide 30, thereby moving rinse fountain 10 and vacuum manifold 1 to theright.

[0053] As shown in FIG. 12, dispense tips 7 are then lowered into thevacuum manifold 11 via linear actuator 26 and the vacuum is turned onfor a programmable time period, thereby drying dispense tips 7. Thiscleaning cycle can be set by the user, via the computer interface, to berepeated as many times as necessary.

[0054] Mounting plate 25 is then raised via linear actuator 26 as shownin FIG. 13. Linear actuator 26 is then moved via linear actuator 21 sothat dispense head 6 is directly above reservoir plate 5, as shown inFIG. 14.

[0055] Mounting plate 25 is then lowered via linear actuator 26 so thattips 7 are dipped into the solution contained in reservoir plate 5, asshown in FIG. 15. While in reservoir plate 5, dispense tips 7 pick upsome of the solution to be dispensed.

[0056] Linear actuator 26 then moves to the first dispense positionshown in FIG. 16A so that dispense head 6 is above slide 4A1. Based onthe earlier positioning data regarding slide 4A1 (see discussion of FIG.4), linear actuator 21 makes minute positioning adjustments to linearactuator 26 and linear actuator 15 makes minute positioning adjustmentsto platform 2 in order to accurately position dispense head 6 over slide4A1 at the first dispense position.

[0057]FIG. 16B shows a top view of a blank slide 4A1. In this preferredembodiment, slide 4A1 has 96 positions 60 arranged in an 8×12 array. Ateach position 60, slide 4A1 is etched so as to be better able to retaina drop of solution deposited at the spot.

[0058] Dispense head 6 is then lowered via linear actuator 26 so thattips 7 are in contact with slide 4A1 at the first dispense position, asshown in FIG. 17A. As tips 7 contact slide 4A1, spots 62 are placed onslide 4A1 via surface tension, as shown in FIG. 17B.

[0059] Dispense head 6 is raised via linear actuator 26, as shown inFIG. 18. Based on the earlier positioning data regarding slide 4A1,linear actuator 21 makes minute positioning adjustments to linearactuator 26 and linear actuator 15 makes minute positioning adjustmentsto platform 2 in order to accurately position dispense head 6 over slide4A1 at the second dispense position.

[0060] Dispense head 6 is then lowered via linear actuator 26 so thattips 7 are in contact with slide 4A1 at the second dispense position, asshown in FIG. 19A. As tips 7 contact slide 4A1, more spots 62 are addedto slide 4A1, as shown in FIG. 19B.

[0061] Dispense head 6 is raised via linear actuator 26, as shown inFIG. 20. Based on the earlier positioning data regarding slide 4A1,linear actuator 21 makes minute positioning adjustments to linearactuator 26 and linear actuator 15 makes minute positioning adjustmentsto platform 2 in order to accurately position dispense head 6 over slide4A1 at the third dispense position.

[0062] Dispense head 6 is then lowered via linear actuator 26 so thattips 7 are in contact with slide 4A1 at the third dispense position, asshown in FIG. 21A. As tips 7 contact slide 4A1, more liquid spots 62,are added to slide 4A1, as shown in FIG. 21B.

[0063] Dispense head 6 is raised via linear actuator 26, as shown inFIG. 22. Based on the earlier positioning data regarding slide 4A1,linear actuator 21 makes minute positioning adjustments to linearactuator 26 and linear actuator 15 makes minute positioning adjustmentsto platform 2 in order to accurately position dispense head 6 over slide4A1 at the fourth dispense position.

[0064] Dispense head 6 is then lowered via linear actuator 26 so thattips 7 are in contact with slide 4A1 at the fourth dispense position, asshown in FIG. 23A. As tips 7 contact slide 4A1, more liquid spots 62 areadded to slide 4A1, as shown in FIG. 23B.

[0065] Dispense head 6 is raised via linear actuator 26, as shown inFIG. 24. Camera 12 and strobe 13 scans slide 4A1 and acquires images andinspects for spot quality. It is at this point that PC control system300 (FIG. 37) identifies slide 4A1 as pass or fail. (Preferred computercontrolled techniques for making this determination are discussed in afollowing section.)

[0066] As shown in FIG. 25, based on the earlier positioning dataregarding slide 4A2 (see discussion regarding FIG. 5), linear actuator21 makes positioning adjustments to linear actuator 26 and linearactuator 15 makes positioning adjustments to platform 2 in order toaccurately position dispense head 6 over slide 4A2 at the first dispenseposition for slide 4A2.

[0067] The four-stage liquid dispense cycle (explained above withrespect to slide 4A1 in discussion regarding FIGS. 17A-24) is repeatedso that at the end of the four-stage cycle, slide 4A2 contains spots 62,as shown in FIG. 26B. At the end of the four stage cycle, dispense head6 is raised via linear actuator 26, as shown in FIG. 26A. Camera 12 andstrobe 13 scans slide 4A2 and acquires images and inspects for spotquality. It is at this point that the control system identifies slide4A2 as pass or fail.

[0068] As shown in FIG. 27, linear actuator 21 moves linear actuator 26so that dispense head 6 is above slide 4A3.

[0069] The sequence outlined in the discussion regarding slides 4A1 and4A2 (depicted in FIGS. 4-26 ) is repeated with regards to slides 4A3 and4A4. To summarize, by utilizing light provided by strobe 13, camera 12will first record the positions of slides 4A3 and 4A4. Then, dispensetips 7 are dipped in sonic cleaner 9. Dispense tips 7 are then rinsed inrinse fountain 10. Then, dispense tips 7 are dried in vacuum manifold11. This cycle is repeated as needed. Then, liquid is picked up bydispense tips 7 when dispense tips 7 are lowered into reservoir plate 5.Then, liquid is spotted onto slide 4A3 by dispense tips 7 in afour-stage liquid dispense cycle. Likewise, liquid is spotted onto slide4A4 in a four-stage liquid dispense cycle so that liquid has beenspotted on both slides, as shown in FIGS. 28B and 28C.

[0070] The process is then repeated for the remaining six slides4A5-4A10 until all the slides on locator plate 4A have been spotted, asshown in FIG. 29.

[0071] Linear actuator 15 then moves platform 2 so that slide 4B1 oflocator plate 4B underneath dispense head 6, as shown in FIG. 30.

[0072] In a similar fashion, dispense tips 7 continue to spot all tenslides on locator plates 4B-4E, until the last slide 4E10 has beenspotted, as shown in FIG. 31.

[0073] After slide 4E10 has been spotted and camera 12 and strobe 13 hasscanned slide 4E10 for spot quality, linear actuator 15 moves platform 2to the position shown in FIG. 32 so an operator can remove locatorplates 4A-4E.

Computer Controlled Pass-Fail Determination Technique

[0074] The computer controlled pass-fail determination techniquedetermines individual spots as pass or fail based on several criteria.For each slide, the camera system scans a region to look for a spot. Ina preferred embodiment the criteria that are applied to that inspectionregion are spot presence, spot size in area, spot location, and spotgeometry. Additional criteria can be added through softwareconfiguration. Each of the criteria can have upper and lower limitsdesignated which define the acceptable values for that particularcriteria. Any value that falls outside of the limits for any criteriaqualifies that spot and slide as failed. The actual inspection valuesare determined by analyzing the grayscale intensity of each pixel. Thetotal number of pixels falling above and below a threshold are talliedto give values for each of the inspection criteria.

Rework Capability

[0075] As explained above, after each slide has been spotted, camera 12and strobe 13 scans the slide and acquires images and inspects for spotquality. It is at this point that the control system identifies theslide as pass or fail. In a preferred embodiment of the presentinvention, an operator monitoring the spotting process via monitor 305(FIG. 37) has the option of correcting a slide that has failed.

[0076] For example, FIG. 29 shows locating plate 4A after all slides4A1-4A10 have been spotted. At this point, an operator can scan locatingplate 4A. A good plate shows up green as in all slides pass. A platewith at least one bad spot on one of the slides shows up red. The usercan then zoom in on the bad slide and the good and bad spots show upgreen and red respectively as pass or fail. From there, the user candecide whether or not to rework the bad spots.

[0077]FIGS. 39A and 39B show where an operator has decided to reworkslide 4A6 that has a spot that has failed quality inspection. Dispensehead 40 is lowered via pneumatic slide 41 so that dispense tip 42 islower than dispense tips 7. Solution from reservoir plate 5 is thendeposited on the slide at the location of the failed spot. If there areother spots that failed, the operator can likewise rework those spots ina similar fashion.

[0078] Although in the description given above regarding the reworkprocess, the operator reworked failed slides after locating plate 4A hadbeen entirely spotted, it is also possible to rework failed slides atother stages during the spotting process. For example, it may bedesirable to wait until all slides 4A1-4E10 on plates 4A-4E have beenspotted (FIG. 31) before reworking them. This allows an operator to befree to do other activities while the initial spotting is taking place.Then, after all slides have been spotted, he can come back and do allthe reworking at one sitting.

[0079] Alternatively, it may be desirable to rework each slideimmediately after it has been spotted, as shown in FIG. 24.

Automatic Rework Capability

[0080] The previous section described a preferred embodiment where anoperator can decide whether or not to rework a spot based on a computerdetermination of pass or fail. In another preferred embodiment therework decision is made automatically by the computer based on whetheror not the spot has passed or failed. In this preferred embodiment, thecomputer makes a determination whether or not a spot has passed orfailed using the computer controlled pass-fail determination techniqueearlier described. If, based on its analysis, the computer determinesthat the spot has failed, the computer will automatically take steps torework the spot. For example, dispense tip 42 will extract solution fromreservoir plate 5. Then, the computer will lower dispense head 40 viapneumatic slide 41 so that dispense tip 42 is lower than dispense tips7, as shown in FIGS. 39A and 39B. Solution from reservoir plate 5 willthen deposited on the slide at the location of the failed spot.

Components of a Preferred Embodiment of the Present Invention Three AxisRobotic Positioning Stage

[0081] In a preferred embodiment, linear actuators 26, 21 and 15 areindustrial grade precision ground ball screw linear actuators, as shownin FIG. 33. These linear actuators are manufactured by Parker Automation( Model #s:404XR and 406XR series). They are each controlled by a smartservomotor (SmartMotor, Model #2320 VRE, manufactured by Animatics, withoffices in Santa Clara Calif.), which is a fully self-contained closedloop servo system. Each of these smart servomotors contains the motor,encoder, amplifier, and controller all in one small package mounted tothe linear actuator. Linear actuator 15 (the x-axis positioning device)has an overall travel distance of 600 mm with an accuracy within+/−0.032 micrometers Linear actuator 21 (the y-axis positioning device)has an overall travel distance of 400 mm with an accuracy within+/−0.032 micrometers. Linear actuator 26 (the z-axis positioning device)has an overall travel distance of 100 mm with an accuracy within +/−micrometers. In this preferred embodiment, this extreme accuracy isneeded to accommodate very small spot size and spacing between spots.The linear actuators have pitches of 5mm per revolution giving apositioning accuracy of 0.032×10⁻⁶ meters.

[0082] Linear actuator 15 controls the positioning of platform 2containing slides 4A1-4E10 along the x-axis of motion making all slidespresentable to the dispense head 6. Linear actuator 21 controls thepositioning of the dispense head along the y-axis of motion making allslides 4A1-4E10, sonic cleaner 9, rinse fountain 10, vacuum manifold 11,and reservoir plate 5 presentable to dispense head 6. Linear actuator 26controls the positioning of dispense head 6 along the z-axis of motionallowing dispense head 6 to be lowered to and raised from all slides4A1-4E10, sonic cleaner 9, rinse fountain 10, vacuum manifold 11, andreservoir plate 5.

Cleaning Station

[0083] Cleaning station 33 consists of sonic cleaner 9, rinsing fountain10, and a drying vacuum manifold 11. In a preferred embodiment, soniccleaner 9 is an ultrasonic cleaner manufactured by Prosonic, Inc. (partno. E0028). Sonic cleaner 9 can contain either a cleaning solution orsimply purified water. Dispense tips 7 are dipped in the sonic cleaner9, where the ultra sonic oscillations of the cleaning solution clean thetips.

[0084] Rinsing fountain 10 and the vacuum manifold 11 are placed on apneumatic slide 30. Pneumatic slide 30 is used to select which operationis to be performed, rinsing or drying. The reason for this slide is sothat both operations can be performed at a single position along they-axis. This allows for both operations without having to increase theoverall travel of linear actuator 26 along the y-axis.

[0085] Rinsing fountain 10 pumps in purified water and drains it out toa waste bin. Dispense tips 7 are dipped in this purified water to rinseaway any debris or cleaning solution that may remain on the tips aftercleaning.

[0086] Drying vacuum manifold 11 is a block with an array of holes in itthat match the array of dispense tips 7. The tips are inserted into theblock, each of these holes are connected to a manifold which isconnected to a vacuum generator and air supply. The vacuum pulls awayany remaining liquid or debris left on the dispense tips after rinsing.

Dispense Head Assemblies

[0087] As shown in FIG. 35B, dispense head 6 is a 4×6 grid Micro QuillHolder (part no. 11946-0) made by Major Precision of Arizona. A 4×6array of primary dispense tips 7 are held in dispense head 6. As shownin FIG. 35A, dispense head 40 is a Micro Quill Holder also made by MajorPrecision. Dispense tip 42 is held in dispense head 40. Dispense tips 7and 42 are spring loaded within the dispense heads 6 and 40.

[0088] As shown in FIGS. 1, 34A and 34B, dispense head 6 is rigidlymounted to mounting plate 25, whereas dispense head 40 is mounted topneumatic slide 41 (manufactured by Robohand, Inc., with offices inMontroe, Conn., part no. MPS1-2). Mounting plate 25 is capable of movingup and down along the z-axis via linear actuator 26. Furthermore,dispense head 40 is capable of independent additional movement up anddown along the z-axis via pneumatic slide 41, as shown in FIGS. 34A-34B.

[0089] During normal operation, such as that depicted in the sequenceillustrated in FIGS. 3-32, dispense head 6 is used to spot slides4A1-4E10. For example, FIG. 34C shows a front view of dispense tips 7 incontact with slide 4A1. Dispense tips 7 will be used to spot slide 4A1at positions 60, as shown in FIG. 34C. Note that when dispense head 6has been selected, dispense head 40 is raised via pneumatic slide 41 sothat dispense tips 42 do not interfere with the spotting process.

[0090] If, however, the operator wishes to spot slide 4A1 at positions61 (FIG. 34C), dispense head 40 will be lowered via pneumatic slide 41so that dispense tips 42 are in contact with slide 4A1 and dispense tips7 are out of the way, as shown in FIG. 34B.

Camera and Lighting

[0091] In a preferred embodiment, camera 12 and strobe 13 are mounted tothe side of linear actuator 26 as shown in FIG. 1. Camera 12 is aself-contained camera with image processing and Ethernet capabilitiesmanufactured by DVT Corporation with offices in Norcross, Ga. (series600 model). Using light provided by strobe 13, camera 12 can snappictures while in dynamic motion, process the image for results, passthe results off to the PC control system, and prepare for the next imageacquisition. The camera uses a 55 mm Telecentric lens which provides theproper field of view and magnification for reading of 2D bar code 62(FIG. 34C) and for image inspection. Strobe light 13 is preferably ModelDL2449, manufactured by Advanced Illumination, with offices inRochester, Vt. In the preferred embodiment, the image acquisition timeis ˜40 ms and the image processing time is ˜50 ms. The system can alsobe equipped with a flouresence device along with the camera for furthergenomic expression analysis.

Vibration Isolated Base

[0092] As shown in FIG. 36, vibration isolated base 80 is provided tominimize any possible affects that high frequency environmentalvibrations might have on the dispensing process. This base is apneumatic system which acts as a shock absorber to the system.

[0093] In a preferred embodiment, the base is manufactured by Newport,Inc. with offices in Irvine, CA, model # CM-225.

PC Based Control System

[0094]FIG. 37 depicts a block diagram of PC control system 300 and othercomponents of a preferred embodiment of the present invention. PCControl System 300 includes CPU 301 with associated memory (RAM 302 andROM 303). It also includes a touch screen monitor/interface 305 thatallows for operator monitoring, intervention and control of the presentinvention. In the preferred embodiment, the computer system is a PCbased computer equipped with an ethernet card and running windowssoftware. The programming is preferably written in VISUAL BASIC. (VISUALBASIC is a federally registered trademark of Microsoft Corp., a DelawareCorporation) PC Control System 300 is equipped with CMS (CentralMonitoring System). The CMS gives PC Control System 300 it's own IP(Internet Protocol) address and ethernet connectivity. This allows forremote monitoring and control via Intranets as well as Internet providedthat the bandwith is available for proper functionality. The software ishighly comfigurable to allow increased flexibility for customers withvarying slide types, slide sizes, slide orientations, spot size, spotspacing and many other variables.

Control of the Components of the Preferred Embodiment of the PresentInvention through the PC Control System

[0095] As previously stated, linear actuators 26, 21 and 15 areindustrial grade precision ground ball screw linear actuators. As shownin FIG. 37, PC control system 300 sends signals to smart servomotors26A, 21A and 15A to control linear actuators 26, 21 and 15,respectively. PC control system 300 controls sonic cleaner 9 and rinsefountain 10. Compressed air source 310 provides compressed air topneumatic slides 30 and 41 via valves 310 controlled by PC controlsystem 300. Vacuum generator 320 provides a vacuum to vacuum manifold 11via valve 317. As previously explained camera 12 and strobe 13 work inconjunction to provide sensory data to PC control system 300. This inputis used to accurately position the dispense heads over the slides toensure optimum spotting and to verify the quality of the spotting as“pass” or “fail” using multiple criteria as to placement at intendedlocation as well as spot size (too big or too small).

Second Preferred Embodiment of the Present Invention

[0096] A second preferred embodiment of the present invention is shownin FIG. 41. In the second preferred embodiment, dispense head 106 isconnected via mounting plate 125 to linear actuator 126 so that dispensetips 107 can be raised and lowered along the z-axis into solution inmicroplate 190. Camera 112 and strobe 113 are rigidly mounted to theside of linear actuator 126 so that they remain stationary with respectto the side of linear actuator 126 along the z-axis. Linear actuator 126is mounted to linear actuator 121 so that it can move along the y-axis.

[0097] Platform 182 is mounted to linear actuator 180 so that it canmove along the x-axis. Locating plate 4A is placed on top of platform182. Platform 102 is mounted to linear actuator 115 so that it can movealong the x-axis. Microplate 190 is place on top of platform 102. Inthis preferred embodiment platform 102 has the capacity to hold tenmicroplates 190.

[0098] Solution in microplate 190 is removed via dispense tips 107.Linear actuator 126 then moves along the y-axis so that dispense tips107 are above locating plate 4A. The solution is then spotted in afashion similar to that described for the earlier preferred embodiments.Camera 112 with strobe 113 is focused so as to permit recording of thedeposition process and functions to permit verification of slideidentification information, permit verification of proper deposition ofsolution on the slides, and to verify slide alignment. As explainedabove, slide image data is transferred via camera 112 to a PC controlsystem where the data is analyzed. The results of the analysis are thenavailable for improving the spotting of the solution onto the slides.For example, spots that have failed to meet the threshold limits can bereworked. Also, the computer can automatically make adjustments to therelative positions of the slides and dispense tips based on the slidealignment analysis. Periodically, during the cycle, the dispense tipsare cleaned in sonic cleaner 109, then rinsed in the rinse fountain anddried in vacuum manifold 111.

[0099] Operation of the First Preferred Embodiment with the SecondPreferred Embodiment

[0100] The first preferred embodiment (described in the sequenceillustrated in FIGS. 3-32) can be used in conjunction with the secondpreferred embodiment to spot slides. For example as shown in FIG. 32,locating plate 4A can be removed from the microarrayer via an operatorafter it has been spotted with a base solution. Locating plate 4A can betransferred to the microarrayer depicted in FIG. 41. It can be placed onplatform 180. DNA from microplate 190 can then be spotted on top of thebase solution spotted already on slides 4A1-4A10.

Use of the Present Invention with Other Microarrayers

[0101] Although the present invention was described as being used withthe preferred microarrayer depicted in the sequence described byreference to FIGS. 3-32, those of ordinary skill in the art willrecognize that it is possible to use camera 12, strobe 13 and a PCcontrol system in conjunction with a variety of microarrayer designs.For example, in the background section of this application, severalmicroarrayers were mentioned. It would be possible to one of ordinaryskill in the art to modify a prior art automatic microarrayer to includecamera 12 and strobe 13. Camera 12 and strobe 13 would then work inconjunction to provide sensory data to PC control system 300, asdescribed above. Also, as explained above, the input would be used toaccurately position the dispense heads over the slides to ensure optimumspotting and to verify the quality of the spotting as “pass” or “fail”.

Modification of Rework Dispense Tips

[0102] The previous embodiments showed one dispense tip 42 extendingdownward from dispense head 40. It was explained how the single dispensetip 42 is used for reworking (correcting) defective spots. It ispossible, however, to modify dispense head 40 so that multiple dispensetips can extend downward from dispense head 40. A preferred embodimentis shown in FIG. 40 in which five dispense tips 42A-E extend down belowdispense head 40. In this preferred embodiment, dispense tips 42A-E areretractably connected to dispense head 40. As shown in FIG. 40, dispensetips 42A-D are retracted inside dispense head 40. The rightmost dispensetip 42E is extended below the other dispense tips and is spotting slide4A1. In a preferred embodiment, dispense tips 42A-E are mounted to apneumatic slides 43.

[0103] An advantage of this embodiment is that each dispense tip 42 canbe configured to dispense a different volume of solution. For example,in a preferred embodiment, dispense tip 42A would dispense 1 nL ofsolution, dispense tip 42B would dispense 2 nL of solution, dispense tip42C would dispense 4 nL of solution, dispense tip 42D would dispense 8nL of solution, and dispense tip 42E would dispense 16 nL of solution.

[0104] After initially spotting the slides as explained above, camera 12and strobe 13 would work in conjunction to provide sensory data to PCcontrol system 300 reporting the quality of the spots. The spots wouldthen be classified as pass or fail. If a spot has failed, the softwarein conjunction with PC control system 300 would determine the amount ofsolution required to correct the failed spot. Then, during the reworkingsequence, the dispense tip that dispenses the most correct volume wouldbe extended down from dispense head 40 and the other dispense tips wouldbe retracted upward inside dispense head 40, as shown in FIG. 40.

Third Preferred Embodiment

[0105] A third preferred embodiment of the present invention is seen byreference to FIG. 42. FIG. 42 shows a perspective view of microarrayer250. In the third preferred embodiment, dispense head 202 is connectedvia mounting plate 213 to linear actuator 214 so that dispense tips 208can be raised and lowered along the z-axis. Camera 215 and strobe 216are also mounted to mounting plate 213 and can be raised and loweredalong the z-axis by linear actuator 214. Linear actuator 214 is mountedto linear actuator 217 so that it can move along the y-axis.

[0106] Platform 218 is mounted to linear actuator 219 so that it canmove along the x-axis. 140 slides 220(1)-220 (140) are mounted on top ofplatform 218. Microplates 204(A -C) and pre-spot slide holder 205 havingpre-spot slide 206 are placed on top of platform 218.

Operation of the Third Preferred Embodiment Removing Solution From theMicroplate

[0107] To remove solution from microplate 204(A), linear actuator 219moves platform 218 along the x-axis and linear actuator 217 moves linearactuator 214 along the y-axis so that dispense head 202 is positionedover microplate 204(A), as shown in FIG. 44. Linear actuator 214 thenlowers dispense head 202 so that dispense tips 208 are dipped into thewells of microplate 204(A). Linear actuator then raises dispense head202. Dispense tips 208 contain solution removed from microplate 204(A).

Pre-Spotting the Solution onto a Pre-Spot Slide

[0108] The third preferred embodiment includes pre-spot slide 206.Pre-spot slide 206 allows excess solution on the dispense tips to beremoved prior to the solution being spotted onto slides 220(1)-220(140). This is achieved by repeatedly pressing the dispense tips ontothe pre-spot slide until the excess solution is removed.

[0109] To pre-spot solution onto pre-spot slide 206, linear actuator 217moves linear actuator 214 along the y-axis so that dispense head 202 ispositioned above pre-spot slide holder 205 having pre-spot slide 206, asshown in FIGS. 45A and 45B. Linear actuator 214 then lowers mountingplate 213. As previously stated, dispense head 202 is mounted tomounting plate 213. Linear actuator lowers mounting plate 213 withdispense head 202 until dispense tips 208 come into contact with thesurface of pre-spot slide 206, as shown in FIG. 45C. The act of dispensetips 208 pressing against the surface of pre-spot slide 206 causesexcess solution on the sides of the dispense tips to come off thedispense tips. The third preferred embodiment is programmed so thatlinear actuator 214 raises and lowers dispense head 202 in a repetitivefashion so that dispense tips 208 are pressed against pre-spot slide 206repetitively until the desired amount of solution is removed. The amountof times dispense tips 208 should be pressed varies depending on theproperties of the solution being deposited and can be modified by theuser by inputting instructions into computer 280 (FIG. 42).

Spotting Solution onto Slides

[0110] After the solution has been pre-spotted onto pre-spot slide 206,the solution can then spotted onto selected microplates 220(1)-220 (140)in a fashion similar to that described above in reference to the firstand second preferred embodiments. For example, to spot solution ontoslide 220(1), linear actuator 219 moves platform 218 along the x-axisand linear actuator 217 moves linear actuator 214 along the y-axis sothat dispense head 202 is positioned over slide 220(1), as shown in FIG.46. The solution is then spotted in a fashion similar to that describedfor the earlier preferred embodiments. Camera 215 with strobe 216 isfocused so as to permit recording of the deposition process andfunctions to permit verification of slide identification information,permit verification of proper deposition of solution on the slides, andto verify slide alignment. As explained above, slide image data istransferred via camera 215 to a PC control system where the data isanalyzed. The results of the analysis are then available for improvingthe spotting of the solution onto the slides. For example, spots thathave failed to meet the threshold limits can be reworked. Also, thecomputer can automatically make adjustments to the relative positions ofthe slides and dispense tips based on the slide alignment analysis.Periodically, during the cycle, the dispense tips are cleaned in soniccleaner 231, then rinsed in the rinse fountain 230 and dried in vacuummanifold 232.

Robotic Loading of Microplates

[0111]FIG. 43 shows robotic plate loader 252 positioned betweenmicroarrayers 250. Microarrayers 250 and robotic plate loader 252 arecontrolled by programmable computer 280.

[0112] As shown in FIG. 43, microplate staging area 254 has 4 microplatestorage racks 255(A)-255(D). Each storage rack is capable of holding upto 15 microplates. Robotic plate loader 252 is capable of removing themicroplates from each of the storage racks. Robotic plate loader 252 canpivot so that arm 257 can be extended towards a desired storage rack.Robotic gripper 256 can then grip the desired plate and remove it fromthe storage rack. The microplate can then be placed on either of theplatforms 218(A) -(B) (as shown) or at pre-staging area 258. Microplatelids 259 can then be removed and placed at lid holding areas 260 and261.

Robotic Plate Loading from Incubator

[0113] By storing microplates in an incubator, the temperature of thesolution inside the microplates can be kept at a controlled level. FIGS.47 and 48 show another preferred embodiment in which microplates storedin incubator 270 are automatically loaded via robotic arm 271 ontoplatform 218. FIG. 48 shows a top view and FIG. 47 shows a perspectiveview of incubator 270.

[0114] In FIG. 48, incubator access door 278 has been raised permittingoutside access to incubator 270. Fifteen microplates are stored insidestorage racks 274(A)-274(D). Robotic pivot arm 275 of robot 272 insideincubator 270 is capable of pivoting about axis 273 so that it can reachstorage racks 274(A)-274(D).

[0115]FIG. 48 shows robotic pivot arm 275 in dotted line positioned sothat gripper 276 is in front of storage rack 274(A). Robotic arm 275 isshown in solid line in a vertical position placing microplate 274(A)(13)on access plate 277.

[0116] In FIG. 47, robotic arm 271 has maneuvered gripper 279 so that itwas able to remove microplate 274(A)(13) from access plate 277, rotateit 90 degrees, and place it on platform 218 as shown.

Automatic Spotting from Multiple Microplates onto Multiple Slides

[0117] In the third preferred embodiment, microarrayer 250 can beprogrammed to remove solution from multiple microplates and spot thesolution onto multiple slides 220 (1-140). FIG. 52A shows a top view ofplatform 218. Microplates 204(A), 204(B) and 204(C) are on platform 218.Microplates 204(D-O) are in storage rack 255(A). Microplates 285(A-O)are in storage rack 255(B). Microplates 286(A-O) are in storage rack255(C) and microplates 287(A-O) are in storage rack 255(D), as shown inFIG. 52AB.

[0118] Robotic plate loader 252 is positioned between platform 218 andstorage racks 255(A-C) and is capable of accessing the positionsoccupied by microplates 204(A-C) and any of the microplate storagepositions in storage racks 255(A-D). Dispense head 202 (see also FIG.42) is capable of being positioned over microplates 204(A-C) viamicroarrayer 250 and is capable of removing solution from microplates204(A-C) and spotting that solution on any of the slides 220(1-140).Robotic plate loader 252 can move microplates 204(A-C) and store themback in storage rack 255(A) and then place other microplates ontoplatform 218. For example, a microarrayer 250 could be programmed toplace microplates 204(A-C) back into storage rack 255(A) and then placemicroplates 285(A), 286(D), and 287(H) onto platform 218.

Flexible Mapping

[0119] In the preferred embodiment, a user of microarrayer 250 (FIG. 42)is able to customize the manner in which spots are deposited on slides220(1-140). FIG. 49A shows a top view of 384 well microplate 204A. Inthe preferred embodiment, microplate 204A is divided into 8 sections204A1-204A8, as shown in FIG. 49B. Each section 204A1-204A8 covers a4×12 array of wells. In one preferred embodiment, a unique solution isdeposited in each section 204A1-204A8.

[0120]FIG. 50 shows a top view of 4×12 pin dispense head 202 next tomicroplate 204(A). Dispense tips P1-P48 are shown as “+” signs and arearranged as shown. The 48 dispense tips on dispense head 202 are spacedto fit into the wells of each 4×12 well section A1-A8 of microplate 204A(FIGS. 51 and 49B). FIG. 49C shows dispense head 202 positioned oversection 204A2 and FIG. 49D shows dispense head 202 positioned oversection 204A5.

[0121]FIG. 53 shows a top view of slide 220(1). Dispense head 202 havingdispense tips P1-P48 dispenses the spots onto slide 220(1). Eachdispense tip P1 -P48 is responsible for dispensing the spots in its own8×8 array. For example, dispense tip PI dispenses all the spots in array283(1) and dispense tip P4 dispenses all the spots in array 283(4),(FIG. 53).

[0122] In the preferred embodiment, a user can program computer 280 sothat dispense head 202 will remove solution from section A1-A8 ofmicroplate 204A and deposit the solution in a customized fashion onslide 220(1).

[0123] For example, in a preferred embodiment a user will use computer280 to control microarrayer 250 and will see on monitor 281 a screenimage similar to that depicted in FIG. 54. At the upper left corner ofthe screen the user inputs the number of microplates for spotting. Inthe example shown, the user has indicated eight plates. Eightmicroplates will provide solution to fill one slide with 3072 spots. Forexample, as shown in FIG. 53 slide 220(1) holds forty-eight 8×8 arraysof spots. Each 8×8 array has 64 spots. Since there are 48 arrays onslide 220(1), there are a total of 48×64=3072 spots on slide 220(1).

[0124] Each microplate has eight 4×12 sections from which dispense head202 can remove solution. Since there are eight microplates from whichdispense head 202 can remove solution and each microplate has eightsections, there are a total of 8×8=64 sections from which solution canbe removed. The total number of sections is displayed in the upper leftcorner of the screen in FIG. 54.

Customized Mapping of the Arrays

[0125] In mapping the forty-eight 8×8 arrays to be deposited on slide220(1) it is only necessary for the user to focus on how one of theforty-eight arrays will appear. This is because each array 283(1-48) isidentical to the other. For example, as shown in FIG. 53, while dispensetip P1 of dispense head 202 (FIG. 50) is depositing solution to positionAl of array 283(1), dispense tip P2 and P28 are depositing solution fromthe same microplate section to positions A1 of array 283(2) and A1 ofarray 283(28), respectively. Moreover, while dispense tip P1 isdepositing solution to position C6 of array 283(1), dispense tip P32 isdepositing solution from the same microplate section to position C6 ofarray 283(32). Therefore, after all 64 spots have been deposited by P1onto array 283(1), dispense tips P2-P48 have simultaneously deposited 64spots to each array 283(2-48) for a total of 3072 spots.

[0126] To map an 8×8 array, the user selects from which plates he wantsto remove solution by entering the plate numbers into boxes 300, asshown in FIG. 54. In the present example the user wants to removesolution from each section of microplates 204(A), 204(B), 204(C),204(D), 285(C), 286(D), 286(H), and 287(B). To select which microplatehe wants to first map, the user mouse clicks on one of the eightmicroplates at the bottom of the screen.

[0127] In FIG. 55, the user has selected microplate 204(A) to map.Microplate 204(A) appears in the upper center of the screen under theheading “Current Plate”. The user then mouse clicks on a section that hewants to map. In the present example the user has mouse clicked onsection 204A1 under the “Current Plate” heading. This causes the 204A1section to darken. The user then mouse clicks on position A1 of spotarray map 301. The numerals 204A1 then appear in the Al position of thearray to indicate that the A1 position has been mapped and section 204A1of microplate 204(A) at the left bottom corner of the screen hasdarkened to indicate that the 204A1 section has been mapped.

[0128] In FIG. 56, the user has mouse clicked on section 204A2 under the“Current Plate” heading. This causes the 204A2 section to darken. Thenthe user mouse clicks on C4 of spot array map 301. The numerals 204A2then appear in the C4 position of the array to indicate that the C4position has been mapped and section 204A2 of microplate 204(A) at theleft bottom corner of the screen has darkened to indicate that the 204A2section has been mapped.

[0129] In a similar fashion, the user can map the rest of microplate204(A) onto spot array map 301. FIG. 57 shows microplate 204(A)completely mapped onto spot array map 301.

[0130] The user can continue to map the rest of the microplates byfollowing the above-described procedures with respect to each of theseven other microplates. For example, FIG. 58 shows spot map array 301completely mapped.

[0131] After having filled in spot map array 301, the user saves hiscustomized mapping and runs microarrayer 250 (FIG. 42). Microarrayer 250will remove solution from microplates 204(A-C) first because they arepositioned on platform 218 so that they can be accessed by dispense head202. After solution has been removed from each of the eight sections ofa microplate, robotic plate loader 252 (FIG. 52A) will return themicroplate to its appropriate storage rack. The robotic plate loaderwill then place the next microplate at the position vacated by theprevious microplate on platform 218. For example, after solution hasbeen removed from microplate 204(A), robotic plate loader 252 willreturn microplate 204(A) to its slot in storage rack 255(A) (FIGS. 52Aand 52B). Robotic plate loader 52 will then place microplate 204(D) atthe spot vacated by microplate 204(A). Microarrayer will continue tospot slide 220(1) until it has been completed spotted, as shown in FIG.53.

[0132] If the user would like to spot more slides with the pattern shownin spot map array 301 in FIG. 58, he inputs into computer 280 the numberof slides he would like to spot. For example, if he would like to spotall of the slides 220(1-140) the user inputs that information into thecomputer and microarrayer 250 will spot slides 220(1- 140). Each slidewill be spotted in the same pattern as the other slide. For example,each slide will be spotted in accordance with the pattern shown in spotmap array 301 in FIG. 58. Similarly, if the user just wants to spotslides 220(1-10), he inputs that information into computer 280 andslides 220(1-10) will be spotted in accordance with the pattern shown inspot map array 301 in FIG. 58.

Spot Inspection Data and Spot Inspection Statistics

[0133] As stated previously, camera 215 with strobe 216 is focused so asto permit recording of the deposition process and spot quality andfunctions to permit verification of slide identification information,permit verification of proper deposition of solution on the slides, andto verify slide alignment. Spot data covering the qualities of each spoton the slides is transferred via camera 215 to a computer 280 systemwhere the data is analyzed and stored. In a preferred embodiment, thisinformation is displayed in table format to the user via monitor 281.

[0134]FIG. 59 shows a preferred spot inspection data table. Informationsuch as spot size and spot offset is reported. Status indicating thespot as either good or bad is reported as well as whether rework wasrequired for the spot.

[0135]FIG. 60 shows a preferred table reporting spot inspectionstatistics. In this table, the user can quickly ascertain the quality ofdispense tips P1-P48 on dispense head 202. For example, by referring toFIG. 60, the user can see that dispense tip P1 deposited 20 spots andthey were all good spots and rework was not required. In contrast, theuser can see that dispense tip P2 also deposited 20 spots but that only15 of them were good. Rework was required for 5 spots. The user can thendetermine that dispense tip P1 is operating effectively, but thatdispense tip P2 might need to be cleaned, repaired or replaced.Preferably, the table shown in FIG. 60 also reports information such asspot diameter statistics for each dispense tip and spot offsetstatistics for each dispense tip.

Exportation of Mapping Information, Spot Inspection Data and SpotInspection Statistics

[0136] In a preferred embodiment, it is possible to export to anothercomputer the data associated with the spotting process. For example, asdiscussed above under the headings “Flexible Mapping” and “CustomizedMapping of the Arrays” the user can customize the mapping process inaccordance with his wishes. The mapping information can then be saved bycomputer 280 and matched with the appropriate slide for later reference.For example, if slide 220(1) was mapped in accordance with spot maparray 301 shown in FIG. 58, then an individual later working with slide220(1) could refer to the saved mapping information and immediately knowthe source microplates for each spot on the slide. Likewise, it is alsopossible to save and export the spot inspection data and spot inspectionstatistics discussed above for later reference.

[0137] In one preferred embodiment, the saved information is exportedvia a computer network to another computer on the computer network. Inanother preferred embodiment, the saved information is saved to acomputer disc and the computer disc is physically transferred to anothercomputer where it can be accessed.

Calibration Target

[0138] Preferably, microarrayer 250 (FIG. 42) includes calibrationtarget 303 (see also FIGS. 52 and 61). As shown in FIG. 61, calibrationtarget 303 includes array 304. Array 304 has a known dimension. Also,the size of the dots of the array is known as well as the distancebetween the dots. The color of calibration target 303 is also known. Tocalibrate camera 215, the user positions it over calibration target 303.Camera 215 is focused onto array 304 and measurements are taken. Themeasurements are compared to the known value. If there arediscrepancies, then adjustments can be made to camera 215 to calibrateit. For example, if there is a discrepancy between the known values andthe size of array 204, or the size of the dots, or the spacing of thedots, then zoom lens of the camera can be adjusted. Or, if there is adiscrepancy between the known color of the calibration target and thereported color (i.e., too light or too dark), the iris or the apertureof the camera can be adjusted.

Extra Sonic Cleaning and Vacuum Stations

[0139] In the preferred embodiment, microarrayer 250 includes two vacuumstations 232 and two sonic cleaning stations 231 (FIG. 52A). By havingtwo sets of stations, the user is better assured of thoroughly cleaningdispense tips P1-P48. Preferably, one sonic station 231 contains waterand the other sonic station 231 contains alcohol.

Parameter Adjustments for Spotting

[0140] In a preferred embodiment, the user is able to adjust parametersrelating to spot quality via computer 280 (FIG. 42). On monitor 281, theuser will see a screen image similar to that shown in FIG. 62. The usercan adjust load parameters and dispense parameters by entering valuesinto boxes 305. The definitions of the adjustable parameters are givenbelow in Table 1: TABLE 1 Load Dwell The amount of time the dispensetips are immersed in the well solution before lifting up. Load OffsetThe distance (in microns) from the bottom of the wells that the dispensetips stop while immersed in the well solution. (Ex: −500 microns meansthat the dispense tips stop 500 microns above the bottom of the well.)Load Speed The speed of the dispense tips as they move up and down overthe microplate wells. Load Repeat The number of times the loading of thedispense tips is repeated as they are positioned over the wells. LoadRepeat The amount of time in the up position before the loading Delay ofthe dispense tips is repeated. Dispense/Load The number of times thedispense tips dispense solution before they are loaded again. DispenseThe amount of time the dispense tips are in contact with Dwell the slidebefore lifting up. Dispense The distance (in microns) the springs of thedispense tips Offset compress before lifting up. Dispense The speed ofthe dispense tips as they move up and down Speed over the slide.Dispense The number of times the dispensing of solution Repeat (i.e.,spotting) is repeated as the dispense tips are positioned over theslide. Disp Repeat The amount of time in the up position before theDelay dispensing of solution is repeated.

[0141] While the above description contains many specifications, thereader should not construe these as limitations on the scope of theinvention, but merely as exemplifications of preferred embodimentsthereof. Those skilled in the art will envision many other possiblevariations are within its scope. For example, although the abovesequence described a dispensing process utilizing slides that have 96dispense positions, those of ordinary skill in the art will recognizethat it is possible to use other slides as well. For example, 384 or1536 position slides could be used. It is also possible to use a blankmicroscope slide with no pre-etched dispense positions. Accordingly, thelocation of the different dispense positions will vary depending on thetype of slide being used. The spacing and orientation of the slide canbe selected by an operator through the maintenance menu on the computerinterface. Also, the previous embodiments disclosed using a strobe lightto illuminate the slide below the camera. One of ordinary skill in theart will recognize that it is possible to illuminate the slide withother light sources besides a strobe light. For example, the slide couldbe illuminated with a camera flash, a constant bright light, or afluorescence device, such as a fluorescent LED. Additionally, the slidecould be illuminated from the side opposite the camera. If afluorescence device is used to illuminate the slide, those of ordinaryskill in the art will recognize that it is possible to add a fluorescentdye to the solution being spotted to achieve more in depthcharacterizations. For example, by using a fluorescent LED and addingfluorescent dye to the solution, greater volume determination can beachieved based on fluorescent intensity of the spot. Also, in thepreferred embodiment, it was mentioned that dispense tips 7 and 42 werequill type dispense tips, it would be obvious to substitute other typesof dispense tips. For example, piezo type dispense tips could also beused. The present invention was taught showing the microarrayerdepositing solution onto a plurality of slides. One of ordinary skill inthe art would recognize that the microarrayer could be used to depositsolution onto receiving surfaces that include a variety of slide typesand surfaces other than slides. For example, the microarrayer coulddeposit solution onto a glass slide, or a slide whose surface has beentreated to enhance the spotting or the experimental results, or slidesthat have porous or semi-porous surfaces, membrane slides, gold coatedslides, slides made of porous silicon, or any other surface that may bebeneficially printed upon. Additionally, while the present invention wastaught using substantially flat slides, it would be obvious to usesurfaces that are slightly to moderately curved to print upon.Accordingly the reader is requested to determine the scope of theinvention by the appended claims and their legal equivalents, and not bythe examples which have been given.

I claim:
 1. A microarrayer for spotting solution onto a receivingsurface, comprising: A. at least one dispense head for spotting thereceiving surface, B. at least one light source capable of illuminatingthe receiving surface, C. at least one camera operating in conjunctionwith said at least one light source, said at least one camera capable ofacquiring and transmitting surface image data, D. a computer programmedto: 1) receive said surface image data from said at least one camera, 2)analyze said surface image data, and 3) generate post analysis databased on said analysis of said surface image data, wherein said postanalysis data comprises information relating to the success or failureof said microarrayer to successfully spot solution onto the receivingsurface. A. an adjustment device for permitting adjustments to be madeto said spotting of solution onto the receiving surface, wherein saidadjustments are based on said post analysis data.
 2. The microarrayer asin claim 1, wherein said receiving surface is a plurality of receivingsurfaces.
 3. The microarrayer as in claim 1, wherein said receivingsurface is a plurality of slides.
 4. The microarrayer as in claim 1,wherein said adjustment device is a reworking device for permitting themicroarrayer operator to rework a spot via the microarrayer based onsaid post analysis data.
 5. The microarrayer as in claim 1, wherein saidadjustment device is a reworking device for permitting said computer torework a spot via the microarrayer based on said post analysis data. 6.The microarrayer as in claim 1, wherein said microarrayer furthercomprises: A. at least one dispense tip for immersion into saidsolution, B. a pre-spot receiving surface, wherein said microarrayerremoves excessive amounts of said solution by pressing said at least onedispense tip against said pre-spot receiving surface.
 7. Themicroarrayer as in claim 1, wherein said microarrayer removes saidsolution from a plurality of microplates, wherein a robot automaticallypositions said microplates within accessible reach of said microarrayer.8. The microarrayer as in claim 7, wherein said plurality of microplatesare stored in storage racks at a storage location, wherein said robot iscapable of retrieving said microplates from said storage racks andreturning said microplates to said storage racks.
 9. The microarrayer asin claim 8, wherein said storage location is an incubator.
 10. Themicroarrayer as in claim 1, further comprising a flexible mapping devicefor allowing the microarrayer operator to customize the pattern in whichsaid dispense head deposits solution onto said receiving surface. 11.The microarrayer as in claim 1, wherein said post analysis date furthercomprises spot inspection data comprising information about thecharacteristics of individual spots.
 12. The microarrayer as in claim 1,wherein said dispense head comprises a plurality of dispense tips,wherein said post analysis data further comprises spot inspectionstatistics comprising information about the performance of said dispensetips.
 13. The microarrayer as in claim 1, wherein said microarrayerfurther comprises a calibration target for calibration of said camera.14. The microarrayer as in claim 1, wherein said microarrayer furthercomprises: A. at least two vacuum stations, and B. at least two soniccleaning stations.
 15. The microarrayer as in claim 1, furthercomprising: A. a plurality of dispense tips, and B. a second adjustmentdevice for allowing the microarrayer operator to adjust the manner inwhich solution is loaded onto said plurality of dispense tips and toadjust the manner in which solution is deposited onto the receivingsurface.
 16. A microarrayer for spotting solution onto a receivingsurface, comprising: A. a dispensing means for spotting the receivingsurface, B. a light source means capable of illuminating the receivingsurface, C. a camera means operating in conjunction with said lightsource means, said camera means capable of acquiring and transmittingsurface image data, D. a computer means programmed to: 1) receive saidsurface image data from said at least one camera means, 2) analyze saidsurface image data, and 3) generate post analysis data based on saidanalysis of said surface image data, wherein said post analysis datacomprises information relating to the success or failure of saidmicroarrayer to successfully spot solution onto the receiving surface.A. an adjustment means for permitting adjustments to be made to saidspotting of solution onto the receiving surface, wherein saidadjustments are based on said post analysis data.
 17. The microarrayeras in claim 16, wherein said receiving surface is a plurality ofreceiving surfaces.
 18. The microarrayer as in claim 16, wherein saidreceiving surface is a plurality of slides.
 19. The microarrayer as inclaim 16, wherein said adjustment means is a reworking means forpermitting the microarrayer operator to rework a spot via themicroarrayer based on said post analysis data.
 20. The microarrayer asin claim 16, wherein said adjustment means is a reworking means forpermitting said computer to rework a spot via the microarrayer based onsaid post analysis data.
 21. The microarrayer as in claim 16, whereinsaid microarrayer further comprises: A. at least one dispense tip forimmersion into said solution, B. a pre-spot receiving surface means,wherein said microarrayer removes excessive amounts of said solution bypressing said at least one dispense tip against said pre-spot receivingsurface means.
 22. The microarrayer as in claim 16, wherein saidmicroarrayer removes said solution from a plurality of microplates,wherein a robot means automatically positions said microplates withinaccessible reach of said microarrayer.
 23. The microarrayer as in claim22, wherein said plurality of microplates are stored in storage racks ata storage location, wherein said robot means is capable of retrievingsaid microplates from said storage racks and returning said microplatesto said storage racks.
 24. The microarrayer as in claim 23, wherein saidstorage location is an incubator.
 25. The microarrayer as in claim 16,further comprising a flexible mapping means for allowing themicroarrayer operator to customize the pattern in which said dispensingmeans deposits solution onto said receiving surface.
 26. Themicroarrayer as in claim 16, wherein said post analysis date furthercomprises spot inspection data comprising information about thecharacteristics of individual spots.
 27. The microarrayer as in claim16, wherein said means comprises a plurality of dispense tips, whereinsaid post analysis data further comprises spot inspection statisticscomprising information about the performance of said dispense tips. 28.The microarrayer as in claim 16, wherein said microarrayer furthercomprises a calibration target means for calibration of said camera. 29.The microarrayer as in claim 16, wherein said microarrayer furthercomprises: A. at least two vacuum stations, and B. at least two soniccleaning stations.
 30. The microarrayer as in claim 16, furthercomprising: A. a plurality of dispense tips, and B. a second adjustmentmeans for allowing the microarrayer operator to adjust the manner inwhich solution is loaded onto said plurality of dispense tips and toadjust the manner in which solution is deposited onto the receivingsurface.